Galloway P G, Mulvihill P, Siedlak S, Mijares M, Kawai M, Padget H, Kim R, Perry G
Institute of Pathology, Case Western Reserve University, Cleveland, Ohio 44106.
Am J Pathol. 1990 Aug;137(2):291-300.
The focus of research on the neurofibrillary pathology (NFP) of Alzheimer disease has been not only to determine the component forming the paired helical filaments but also to determine whether they result from abnormal processes affecting a single protein. Therefore, although these studies have lead to controversy concerning the respective contribution of components of microtubules and neurofilaments, there has been essentially no consideration of whether other cytoskeletal systems might also be involved and of what are the common features for the incorporated components. Particularly relevant to this issue is our finding that several antisera raised to either skeletal or smooth muscle tropomyosin, a microfilament component, intensely recognize NFP. These antibodies continue to recognize NFP after affinity purification to tropomyosin or paired helical filament fractions. We show that the antibodies do not recognize NFP due to cross-reactivity with the previously identified NFP components related to neurofilaments and microtubules, tau, and MAP2, or neurofilament proteins because the antisera did not recognize these proteins on immunoblots or were not adsorbable by the proteins. Ultrastructural analysis of the immunoreaction showed that tropomyosin-related epitopes were clustered rather than uniformly distributed along paired helical and straight filaments. Although the distribution suggests that tropomyosin is an NFP-associated protein, its retention by paired helical and straight filaments after detergent extraction indicates that it is an integral component strongly and specifically associated with the filaments characteristic of NFP. These findings indicate that NFP involves the three primary neuronal cytoskeletal filament systems, microtubules, neurofilaments, as well as microfilaments, and therefore that NFP probably results from the reorganization of these normal filaments that interact to comprise the cytomatrix and may continue this interaction under the pathologic condition of Alzheimer's disease to generate novel, abnormal polymers.
阿尔茨海默病神经原纤维病理学(NFP)的研究重点不仅在于确定构成双螺旋丝的成分,还在于确定它们是否源于影响单一蛋白质的异常过程。因此,尽管这些研究引发了关于微管和神经丝成分各自贡献的争议,但基本上没有考虑其他细胞骨架系统是否也会涉及,以及纳入成分的共同特征是什么。与这个问题特别相关的是我们的发现,即几种针对骨骼肌或平滑肌原肌球蛋白(一种微丝成分)产生的抗血清能强烈识别NFP。这些抗体在亲和纯化至原肌球蛋白或双螺旋丝组分后仍能识别NFP。我们表明,这些抗体识别NFP并非由于与先前鉴定的与神经丝和微管相关的NFP成分、tau和MAP2或神经丝蛋白发生交叉反应,因为抗血清在免疫印迹上不识别这些蛋白,或者不能被这些蛋白吸附。免疫反应的超微结构分析表明,原肌球蛋白相关表位沿双螺旋丝和直丝呈簇状分布而非均匀分布。尽管这种分布表明原肌球蛋白是一种与NFP相关的蛋白,但去污剂提取后它在双螺旋丝和直丝上的保留表明它是一种与NFP特征性丝紧密且特异性结合的整合成分。这些发现表明NFP涉及三种主要的神经元细胞骨架丝系统,即微管、神经丝以及微丝,因此NFP可能源于这些正常丝的重组,这些丝相互作用构成细胞基质,并可能在阿尔茨海默病的病理状态下继续这种相互作用以产生新的异常聚合物。